Loading...
Microfluidic system evaluation for the semi-automatic detection of MOG-IgG in serum samples
Author(s)
Khemthongcharoen, Numfon
Uawithya, Panapat
Yookong, Nutthapon
Chanasakulniyom, Mayuree
Jeamsaksiri, Wutthinan
Sripumkhai, Witsaroot
Pattamang, Pattaraluck
Juntasaro, Ekachai
Thienthong, Therdthai
Promptmas, Chamras
Date Issued
December 1, 2021
Type
Article
Abstract
Myelin oligodendrocyte glycoprotein reactive immunoglobulin G antibody (MOG-IgG) is in patients with central nervous system demyelination. Reliability of the conventional detection method relies on technician skills and pipetting error accumulation. This work develops a microfluidic system for semi-automatic MOG-IgG detection using cell-based immunofluorescence (IF) assay. The polydimethylsiloxane (PDMS) microfluidic was modified by poly-L-lysine to enhance the adhesion of Human embryonic kidney (HEK) cell. The untransfected and GFP-MOG transfected HEK cells were cultured, fixed, and stained in the microfluidic with the feeding reagents regulated by a syringe pump. Cell characterization, limit of detection (LOD), and turnaround time of the IF assay operation in microfluidic were compared to those in standard microplate. In microfluidic, cell-clumping formation can be avoided and thus signal variations that are caused by cell overlapping can be significantly reduced. LOD of MOG-IgG detection in the microfluidic is at least 2.5 times better than that in the microplate. Signal intensities of the IF staining for 1 h in microfluidic are comparable to those stained for overnight in the standard microplate. By integration with a serial dilution microfluidic, the optimal cutoff titer for MOG-IgG positivity in the patient samples was determined by Receiver operating characteristic curve (ROC) analysis.
Citation
Sensing and Bio Sensing Research, 34, 2021